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Methods Mol Biol. 2016;1354:61-70. doi: 10.1007/978-1-4939-3046-3_5.

HIV-1 Reverse Transcriptase-Based Assay to Determine Cellular dNTP Concentrations.

Author information

1
Center for Drug Discovery, Emory Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, 1760 Haygood Drive, Health Sciences Research Building, Atlanta, GA, 30322, USA.
2
Center for Drug Discovery, Emory Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, 1760 Haygood Drive, Health Sciences Research Building, Atlanta, GA, 30322, USA. baek.kim@emory.edu.
3
College of Pharmacy, Kyung-Hee University, Seoul, South Korea. baek.kim@emory.edu.

Abstract

Deoxynucleoside triphosphates (dNTPs) are the building blocks of DNA and their biosynthesis is tightly regulated in the cell. HPLC-MS and enzyme-based methods are currently employed to determine dNTP concentrations from cellular extracts. Here, we describe a highly efficient, HIV-1 reverse transcriptase (RT)-based assay to quantitate dNTP concentrations. The assay is based on the ability of HIV-1 RT to function at very low dNTP concentrations, thus providing for the high sensitivity of detection.

KEYWORDS:

Deoxynucleoside triphosphates; HIV-1; Reverse transcriptase; dNTP concentration

PMID:
26714705
PMCID:
PMC4839283
DOI:
10.1007/978-1-4939-3046-3_5
[Indexed for MEDLINE]
Free PMC Article

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